Use for dendropanax morbifera extract for adjusting 15- hydroxyprostaglandin dehydrogenase and pge 2 activity
- Summary
- Abstract
- Description
- Claims
- Application Information
AI Technical Summary
Benefits of technology
Problems solved by technology
Method used
Image
Examples
example 1
Preparation of Dendropanax morbifera Leaf Extract
[0115](1) Preparation of Extract
[0116]Collected D. morbifera leaves were dried in the shade at room temperature. Dried leaves were extracted with methanol for 3 times. The methanol extracts described above were filtered through Whatman No. 1 filter paper, and combined methanol extracts above described were concentrated in vacuum using rotary evaporator.
[0117]Methanol extracts were suspended in water and then fractionated according to the polarity by hexane, diethyl ether, ethyl acetate, n-butanol and water. Each of the extracts was concentrated using a rotary evaporator. This process is illustrated in FIG. 4.
[0118](2) Fractionation and Separation Process
[0119]Using gradient solvent system [CHCl3 (0.1% HCOOH) 100% through CHCl3 (0.1% HCOOH)-MeOH (0.1% HCOOH) / 55:45 for 120 min], 10 fractions were provided (DEE01-DEE10) by conducting the activity-guided fractionation under 254 and 280 nm with Isolera Flash Purification system (BIOTAGE, S...
example 2
Identification of 15-PGDH Expression
[0124](1) Expression and Purification of 15-PGDH
[0125]Escherichia coli BL-21 DE3 cells were transformed by a PGEX-2T expression vector including recombinant 15-PGDH between BamH I and EcoR I as illustrated in FIG. 5.
[0126]The cells were grown in 500 mL medium containing 50 μg / mL ampicillin at 37° C. and 220 rpm until the O.D. at 600 nm reached 0.6. IPTG (isoprophyl β-D-thiogalactoside, 1M stock solution) was added and the cells were grown for 12 hours at 25° C.
[0127]Later, the cells were centrifuged at 4000×g, 4° C. for 30 minutes and collected and the cell pellet was resuspended in 20 mL cold lysis buffer (1× PBS buffer pH 7.4 containing 1 mM EDTA and 0.1 mM DTT) and disrupted by ultrasonication.
[0128]The supernatant was slowly applied to glutathione-sepharose 4B column and equilibrated at 4° C. using lysis buffer. The column described above was rinsed with lysis buffer until O.D. at 280 nm was measured lower than 0.005. Then, 15-PGDH was eluted ...
example 3
Analysis of Cell Viability
[0137](1) Cell Culture
[0138]The HaCaT cells, human keratinocyte cell line, were cultured using DMEM (Dulbecco's modified Eagle's media) in 5% CO2 humidified incubator at 37° C.
[0139]The A549 cells originated from adenocarcinoma-like human alveolar type II were cultured in RPMI medium, and 5% CO2 humidified incubator at 37° C. And then, LNCaP•FGC (androgen-dependent prostate cancer) cells were cultured in RPMI medium and 5% CO2 humidified incubator at 37° C. All culture media were supplemented with heat-inactivated FBS and 100 μg / mL penicillin
[0140](2) Cytotoxicity Test
[0141]The cytotoxicity was determined by MTT assay (Mosmann, 1983).
[0142]The HaCaT cells (1×104 / mL) and LNCaP•FGC cells (4×104 / mL) were seeded per 96 μL of DMEM medium on 96-well plate. After overnight culture, drugs were treated for 72 hours and cultured in 10 μL of MTT (5 mg / mL stock solution) for 4 hours. Then, the medium was removed and formazan was dissolved by adding 150 μL of DMSO. Usin...
PUM
Login to view more Abstract
Description
Claims
Application Information
Login to view more - R&D Engineer
- R&D Manager
- IP Professional
- Industry Leading Data Capabilities
- Powerful AI technology
- Patent DNA Extraction
Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic.
© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap